<i>U7</i> snRNA mutations in <i>Drosophila</i> block histone pre-mRNA processing and disrupt oogenesis

Godfrey, Ashley C.; Kupsco, Jeremy; Burch, Brandon D.; Zimmerman, Ryan M.; Domiński, Zbigniew; Marzluff, William F.; Duronio, Robert J.

doi:10.1261/rna.2270406

Cited by 41 publications

(73 citation statements)

References 58 publications

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“…Because of maternal loading of U7 snRNA, the third instar is the earliest time during development when U7 snRNA is depleted and the U7 mutant phenotype occurs (Godfrey et al, 2006). We confirmed the disruption of the U7 snRNP by assaying for Lsm11, a U7 snRNP-specific protein that no longer accumulates in the HLB in U7 mutant cells ( Figure 5, B and D).…”

Section: Mpm-2 Foci Assemble In the Absence Of The Histone Pre-mrna Pmentioning

confidence: 52%

“…Slbp 15 (Sullivan et al, 2001), stg 4 (Edgar and O'Farrell, 1989), cyclin E AR95 (Knoblich et al, 1994), U7 14 (Godfrey et al, 2006), Df(3R)stg AR2 (Lehman et al, 1999), the histone locus deletion Df(2L)Ds6 (Moore et al, 1983), cyclin E hsp70 , UAS:YFP-Lsm11 (Liu et al, 2006), and daGAL4 (Wodarz et al, 1995) were all described previously. cyclin E mutant embryos were unambiguously identified using a CyO P [wg-lacZ] balancer chromosome.…”

Section: Drosophila Stocksmentioning

confidence: 99%

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Developmental and Cell Cycle Regulation of theDrosophilaHistone Locus Body

White¹,

Leslie

Calvi

et al. 2007

MBoC

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127

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Cyclin E/Cdk2 is necessary for replication-dependent histone mRNA biosynthesis, but how it controls this process in early development is unknown. We show that in Drosophila embryos the MPM-2 monoclonal antibody, raised against a phosphoepitope from human mitotic cells, detects Cyclin E/Cdk2-dependent nuclear foci that colocalize with nascent histone transcripts. These foci are coincident with the histone locus body (HLB), a Cajal body-like nuclear structure associated with the histone locus and enriched in histone pre-mRNA processing factors such as Lsm11, a core component of the U7 small nuclear ribonucleoprotein. Using MPM-2 and anti-Lsm11 antibodies, we demonstrate that the HLB is absent in the early embryo and occurs when zygotic histone transcription begins during nuclear cycle 11. Whereas the HLB is found in all cells after its formation, MPM-2 labels the HLB only in cells with active Cyclin E/Cdk2. MPM-2 and Lsm11 foci are present in embryos lacking the histone locus, and MPM-2 foci are present in U7 mutants, which cannot correctly process histone pre-mRNA. These data indicate that MPM-2 recognizes a Cdk2-regulated protein that assembles into the HLB independently of histone mRNA biosynthesis. HLB foci are present in histone deletion embryos, although the MPM-2 foci are smaller, and some Lsm11 foci are not associated with MPM-2 foci, suggesting that the histone locus is important for HLB integrity. INTRODUCTIONCell cycle-regulated histone protein biosynthesis is controlled primarily through the regulation of histone mRNA abundance, which in cultured mammalian cells increases 35-fold at the G1-S transition (Breindl and Gallwitz, 1973;Borun et al., 1975;Detke et al., 1979;Parker and Fitschen, 1980;DeLisle et al., 1983;Heintz et al., 1983;Harris et al., 1991). This rapid rise in mRNA is achieved by increases in the rate of transcription initiation and pre-mRNA processing as cells enter S phase, followed by rapid degradation of histone mRNA at the end of S phase . How these various aspects of histone mRNA metabolism are linked to other events that drive progression through the cell cycle by regulating the activity of the cyclindependent kinases (Cdks) remains incompletely understood.In animal cells Cyclin E/Cdk2 promotes the G1-to-S transition in part by phosphorylating proteins that mediate changes in gene expression associated with the onset of DNA replication (e.g., pRb; Du and Pogoriler, 2006). These include proteins that regulate histone expression. For example, human NPAT and human HIRA are Cyclin E/Cdk2 substrates that act to stimulate and repress, respectively, histone gene transcription in cell culture experiments (Ma et al., 2000;Zhao et al., 2000;Hall et al., 2001;Nelson et al., 2002;Miele et al., 2005). How the activity of such factors is modulated by Cyclin E/Cdk2 and integrated into cell cycle-regulated histone gene expression in vivo is not known.Cyclin E/Cdk2 may also regulate features of histone mRNA biosynthesis other than transcription, such as premRNA processing. Rather than being polyaden...

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Section: Mpm-2 Foci Assemble In the Absence Of The Histone Pre-mrna Pmentioning

confidence: 52%

Section: Drosophila Stocksmentioning

confidence: 99%

Developmental and Cell Cycle Regulation of theDrosophilaHistone Locus Body

White¹,

Leslie

Calvi

et al. 2007

MBoC

Self Cite

127

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“…118 In Drosophila the maximal requirement for histone mRNA synthesis is likely to be during oogenesis when large amounts of maternal histone mRNA and protein are deposited into the egg. Several mutations that affect the HLB and histone pre-mRNA processing result in female sterility, 32,72,105,107 suggesting that the HLB may be particularly important during oogenesis, either in proper development of the egg 107 or in production of sufficient maternal histone mRNA for the embryo. 105,108 Interestingly, one well-studied organism, C. elegans, apparently lacks an HLB, although their histone mRNAs end in a stem loop and associate with SLBP.…”

Section: Discussionmentioning

confidence: 99%

“…105 Sufficient polyadenylated histone mRNA is produced in the absence of processing factors (e.g., U7 snRNP) to allow much of development to occur in Drosophila. 106,107 We can detect both poly AC histone RNA and nascent read-through unprocessed RNAs by nuclease protection and in situ hybridization assays, allowing us to quantitatively determine the efficiency of histone pre-mRNA processing in various tissues. 72,108 Concentration of U7 snRNP and FLASH in the HLB appears to be particularly important for histone mRNA biosynthesis.…”

Section: Hlb Functionmentioning

confidence: 99%

Coordinating cell cycle-regulated histone gene expression through assembly and function of the Histone Locus Body

2017

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Metazoan replication-dependent (RD) histone genes encode the only known cellular mRNAs that are not polyadenylated. These mRNAs end instead in a conserved stem-loop, which is formed by an endonucleolytic cleavage of the pre-mRNA. The genes for all 5 histone proteins are clustered in all metazoans and coordinately regulated with high levels of expression during S phase. Production of histone mRNAs occurs in a nuclear body called the Histone Locus Body (HLB), a subdomain of the nucleus defined by a concentration of factors necessary for histone gene transcription and premRNA processing. These factors include the scaffolding protein NPAT, essential for histone gene transcription, and FLASH and U7 snRNP, both essential for histone pre-mRNA processing. Histone gene expression is activated by Cyclin E/Cdk2-mediated phosphorylation of NPAT at the G1-S transition. The concentration of factors within the HLB couples transcription with pre-mRNA processing, enhancing the efficiency of histone mRNA biosynthesis. KEYWORDSCell cycle; Drosophila; histone genes; mRNA processing; nuclear bodyProper utilization of genetic information in eukaryotes requires extensive three-dimensional organization of the genome and its associated proteins within the nucleus. The basic unit of genome organization is the nucleosome, an octamer of two molecules of each of the four core histone proteins (H2A, H2B, H3 and H4) that packages »147 base pairs of DNA. In mammalian cells, approximately 4£10 8 molcules of each core histone must be synthesized during S phase of the cell cycle to package the newly replicated DNA. Defects in this process result in genome instability that can contribute to human pathologies like cancer. Histone protein synthesis results from a large increase in production at the beginning of S phase of equal amounts of mRNAs encoding the four core nucleosomal histones, as well as histone H1. This highly coordinated gene expression event is performed by a set of transcription and pre-mRNA processing factors unique to replication dependent (RD) histone mRNA biosynthesis that assemble into a nuclear body tightly associated with RD histone genes called the histone locus body (HLB). HLB formation involves a combination of ordered and stochastic assembly steps, and cell cycle regulated changes in the HLB occur to activate histone gene expression during S phase. Here we describe the history of how the HLB was discovered followed by a discussion of HLB assembly mechanism and how the HLB functions in RD histone mRNA biosynthesis.

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“…Many insertions thought initially to be within intergenic regions have subsequently been mapped to genes as the annotation improved. Many such lines have been used to functionally characterize novel genes, promoters, piRNA clusters, and small RNA genes (for example, Brennecke et al 2003Brennecke et al , 2007Godfrey et al 2006). The GDP recognizes that lines added to the collection on the basis of the above criteria are not equally valuable.…”

Section: Strain Selectionmentioning

confidence: 99%

The Drosophila Gene Disruption Project: Progress Using Transposons With Distinctive Site Specificities

et al. 2011

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The Drosophila Gene Disruption Project (GDP) has created a public collection of mutant strains containing single transposon insertions associated with different genes. These strains often disrupt gene function directly, allow production of new alleles, and have many other applications for analyzing gene function. Here we describe the addition of 7600 new strains, which were selected from .140,000 additional P or piggyBac element integrations and 12,500 newly generated insertions of the Minos transposon. These additions nearly double the size of the collection and increase the number of tagged genes to at least 9440, approximately two-thirds of all annotated protein-coding genes. We also compare the site specificity of the three major transposons used in the project. All three elements insert only rarely within many Polycomb-regulated regions, a property that may contribute to the origin of "transposon-free regions" (TFRs) in metazoan genomes. Within other genomic regions, Minos transposes essentially at random, whereas P or piggyBac elements display distinctive hotspots and coldspots. P elements, as previously shown, have a strong preference for promoters. In contrast, piggyBac site selectivity suggests that it has evolved to reduce deleterious and increase adaptive changes in host gene expression. The propensity of Minos to integrate broadly makes possible a hybrid finishing strategy for the project that will bring .95% of Drosophila genes under experimental control within their native genomic contexts.

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U7 snRNA mutations in Drosophila block histone pre-mRNA processing and disrupt oogenesis

Cited by 41 publications

References 58 publications

Developmental and Cell Cycle Regulation of theDrosophilaHistone Locus Body

Developmental and Cell Cycle Regulation of theDrosophilaHistone Locus Body

Coordinating cell cycle-regulated histone gene expression through assembly and function of the Histone Locus Body

The Drosophila Gene Disruption Project: Progress Using Transposons With Distinctive Site Specificities

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